Large-scale production of human blastoids amenable to modeling blastocyst development and maternal-fetal cross talk

Leqian Yu; Deirdre Logsdon; Carlos A Pinzon-Arteaga; Jialei Duan; Toshihiko Ezashi; Yulei Wei; Ana Elisa Ribeiro Orsi; Seiya Oura; Lizhong Liu; Lei Wang; Kun Liu; Xiaoyun Ding; Linfeng Zhan; Junfei Zhang; Asrafun Nahar; Caitlen Stobbe; Mandy Katz-Jaffe; William B Schoolcraft; Tao Tan; Gary C Hon; Ye Yuan; Jun Wu

doi:10.1016/j.stem.2023.08.002

Large-scale production of human blastoids amenable to modeling blastocyst development and maternal-fetal cross talk

Cell Stem Cell. 2023 Sep 7;30(9):1246-1261.e9. doi: 10.1016/j.stem.2023.08.002.

Authors

Leqian Yu¹, Deirdre Logsdon², Carlos A Pinzon-Arteaga³, Jialei Duan⁴, Toshihiko Ezashi², Yulei Wei⁵, Ana Elisa Ribeiro Orsi⁶, Seiya Oura³, Lizhong Liu³, Lei Wang⁴, Kun Liu⁷, Xiaoyun Ding⁸, Linfeng Zhan⁹, Junfei Zhang¹⁰, Asrafun Nahar², Caitlen Stobbe², Mandy Katz-Jaffe², William B Schoolcraft², Tao Tan⁹, Gary C Hon¹¹, Ye Yuan¹², Jun Wu¹³

Affiliations

¹ Department of Molecular Biology, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA; State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China; Institute for Stem Cell and Regeneration, Chinese Academy of Sciences, Beijing 100101, China; Beijing Institute for Stem Cell and Regenerative Medicine, Beijing 100101, China.
² Colorado Center for Reproductive Medicine, Lone Tree, CO 80124, USA.
³ Department of Molecular Biology, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA.
⁴ Cecil H. and Ida Green Center for Reproductive Biology Sciences, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA.
⁵ Department of Molecular Biology, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA; State Key Laboratory of Animal Biotech Breeding, College of Biological Sciences, China, Agricultural University, Beijing, 100193, China.
⁶ Department of Molecular Biology, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA; Department of Genetics and Evolutionary Biology, Institute of Biosciences, University of São Paulo, São Paulo 05508-090, São Paulo, Brazil.
⁷ Colorado Center for Reproductive Medicine, Lone Tree, CO 80124, USA; Paul M. Rady Department of Mechanical Engineering, University of Colorado, Boulder, CO 80309, USA.
⁸ Paul M. Rady Department of Mechanical Engineering, University of Colorado, Boulder, CO 80309, USA.
⁹ State Key Laboratory of Primate Biomedical Research Institute of Primate Translational Medicine, Kunming University of Science and Technology, Kunming 650500, Yunnan, China; Yunan Key Laboratory of Primate Biomedical Research, Kunming 650500, Yunnan, China.
¹⁰ State Key Laboratory of Animal Biotech Breeding, College of Biological Sciences, China, Agricultural University, Beijing, 100193, China.
¹¹ Cecil H. and Ida Green Center for Reproductive Biology Sciences, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA; Department of Obstetrics and Gynecology, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA; Department of Bioinformatics, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA. Electronic address: gary.hon@utsouthwestern.edu.
¹² Colorado Center for Reproductive Medicine, Lone Tree, CO 80124, USA. Electronic address: yyuan@ccrmivf.com.
¹³ Department of Molecular Biology, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA; Cecil H. and Ida Green Center for Reproductive Biology Sciences, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA; Hamon Center for Regenerative Science and Medicine, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA. Electronic address: jun2.wu@utsouthwestern.edu.

PMID: 37683605
DOI: 10.1016/j.stem.2023.08.002

Abstract

Recent advances in human blastoids have opened new avenues for modeling early human development and implantation. One limitation of our first protocol for human blastoid generation was relatively low efficiency. We now report an optimized protocol for the efficient generation of large quantities of high-fidelity human blastoids from naive pluripotent stem cells. This enabled proteomics analysis that identified phosphosite-specific signatures potentially involved in the derivation and/or maintenance of the signaling states in human blastoids. Additionally, we uncovered endometrial stromal effects in promoting trophoblast cell survival, proliferation, and syncytialization during co-culture with blastoids and blastocysts. Side-by-side single-cell RNA sequencing revealed similarities and differences in transcriptome profiles between pre-implantation blastoids and blastocysts, as well as post-implantation cultures, and uncovered a population resembling early migratory trophoblasts during co-culture with endometrial stromal cells. Our optimized protocol will facilitate broader use of human blastoids as an accessible, perturbable, scalable, and tractable model for human blastocysts.

Keywords: endometrial stromal cells; human blastocyst-like structures; human blastocysts; human blastoids; human integrated stem cell embryo model; human peri-implantation development; naive human embryonic stem cells; naive human induced pluripotent stem cells; syncytiotrophoblast.

Publication types

Research Support, Non-U.S. Gov't
Research Support, N.I.H., Extramural

MeSH terms

Blastocyst
Cell Survival
Embryo Implantation*
Humans
Signal Transduction*
Trophoblasts

Grants and funding

S10 OD028630/OD/NIH HHS/United States